Type iii nkt cells and related compositions and methods

Abstract

The present disclosure relates to type III natural killer T (NKT) cells (e.g., CD3+CD56+ type III NKT cells), pharmaceutical compositions, and methods of preparation and use thereof, in particular use of them as therapeutic agents for the treatment various cancers. Modified type III NKT cells, e.g., to express a chimeric antigen receptor (CAR), a T cell receptor (TCR), a T cell receptor mimic antibody (TCRm), or a combination thereof, pharmaceutical compositions, and methods of preparation and use thereof, are also disclosed.

1 . A method of treating or preventing a cancer, comprising administering to a subject in need thereof a therapeutically effective amount of isolated CD3 + CD56 +  type III natural killer T cells, wherein the cancer is a solid tumor or a hematological malignancy; and wherein optionally the cancer is resistant or refractory to treatment in the absence of the cells. 
     
     
         2 . The method of  claim 1 , wherein the cancer is selected from a B-cell malignancy, leukemia, lymphoma, myeloma, or melanoma; acute myeloid leukemia (AML), B-cell precursor acute lymphoblastic leukemia (B-ALL), non-Hodgkin lymphoma (NHL), chronic lymphocytic leukemia (CLL), Ewing sarcoma, osteosarcoma, fibrosarcoma, rhabdomyosarcoma, mantle cell carcinoma, breast cancer or breast adenocarcinoma, lung adenocarcinoma, ovarian cancer, multiple myeloma, glioblastoma, hepatocellular cancer or hepatocellular carcinoma, neuroblastoma, metastatic melanoma, synovial sarcoma, bladder cancer, esophageal cancer, head and neck cancer, non-small cell lung cancer, prostate cancer, T cell lymphoma, or colon adenocarcinoma; and wherein the cancer is resistant or refractory to treatment in the absence of the cells. 
     
     
         3 . The method of  claim 1 , wherein the cells are CD3 + CD4 + CD56 + , CD3 + CD8 + CD56 + , or CD3 + CD4 − CD8 − CD56 +  cells, each optionally isolated from a biological sample of the subject or a donor. 
     
     
         4 . The method of  claim 3 , wherein the biological sample comprises blood, bone marrow, lymph node tissue, spleen tissue, tumor tissue, one or more induced pluripotent stem cells, and/or one or more peripheral blood mononuclear cells; and wherein optionally the blood comprises peripheral blood and/or umbilical cord blood. 
     
     
         5 . The method of  claim 1 , wherein the cells are isolated from one or more peripheral blood mononuclear cells. 
     
     
         6 . The method of  claim 1 , wherein the cells are modified to express a chimeric antigen receptor (CAR) and comprise one or more polynucleotides encoding the CAR. 
     
     
         7 . The method of  claim 6 , wherein the CAR comprises at least an antigen binding domain, a transmembrane domain, and an intracellular signaling domain. 
     
     
         8 . The method of  claim 7 , wherein the antigen binding domain is capable of binding to CD19, IGF1R, ROR1, BCMA, CD123, CD33, CD38, CD138, CLL-1, LILRB4, GD2, CD20, CD22, CD30, MSLN, EGFRvIII, EGFR, HER2, MUC1, EPCAM, PSMA, SLAMF7, GPC3, or PD-L1. 
     
     
         9 . The method of  claim 7 , wherein the antigen binding domain is capable of binding to CD19, and the cancer is a B-cell malignancy, e.g., B-cell precursor acute lymphoblastic leukemia (B-ALL), non-Hodgkin lymphoma (NHL), or chronic lymphocytic leukemia (CLL); or wherein the antigen binding domain is capable of binding to ROR1, and the cancer is Ewing sarcoma, osteosarcoma, fibrosarcoma, rhabdomyosarcoma, chronic lymphocytic leukemia, mantle cell carcinoma, breast cancer, lung adenocarcinoma, melanoma, neuroblastoma, or ovarian cancer; or wherein the antigen binding domain is capable of binding to BCMA, and the cancer is multiple myeloma; or wherein the antigen binding domain is capable of binding to CD123, CD33, CD38, CD138, CLL-1, or LILRB4, and the cancer is AML; or wherein the antigen binding domain is capable of binding to EGFRvIII or EGFR, and the cancer is glioblastoma; wherein the antigen binding domain is capable of binding to GPC3, and the cancer is hepatocellular carcinoma. 
     
     
         10 . The method of  claim 7 , wherein the antigen binding domain comprises an antibody or an antigen binding fragment thereof, or a non-antibody protein scaffold; wherein the antibody is a monoclonal antibody, a polyclonal antibody, a synthetic antibody, a human antibody, a humanized antibody, or a single domain antibody; wherein the antigen binding fragment comprises a single chain variable fragment (scFv); wherein the intracellular signaling domain comprises a functional signaling domain of at least one stimulatory molecule. 
     
     
         11 . The method of  claim 10 , wherein the at least one stimulatory molecule comprises a zeta chain associated with a T cell receptor complex or a CD3 zeta chain; wherein the intracellular signaling domain further comprises a functional signaling domain of at least one costimulatory molecule; and wherein optionally the at least one costimulatory molecule comprises 4-1BB, CD28, CD27, CD134 (OX40), ICOS, DAP10, or DAP12. 
     
     
         12 . The method of  claim 1 , wherein the cells are modified to express a T cell receptor (TCR) and comprise one or more polynucleotides encoding the TCR; and wherein the TCR comprises at least an alpha chain and a beta chain. 
     
     
         13 . The method of  claim 12 , wherein the alpha chain and/or the beta chain is capable of binding to an antigen, and wherein the antigen is an intracellular antigen, NY-ESO-1, WT1, or MAGE-A3. 
     
     
         14 . The method of  claim 13 , wherein the antigen is NY-ESO-1 and the cancer is neuroblastoma, myeloma, metastatic melanoma, synovial sarcoma, bladder cancer, esophageal cancer, hepatocellular cancer, head and neck cancer, non-small cell lung cancer, ovarian cancer, prostate cancer, or breast cancer; or wherein antigen is WT1 and the cancer is AML. 
     
     
         15 . The method of  claim 1 , wherein the cells are modified to express a T cell receptor mimic antibody (TCRm) and comprise one or more polynucleotides encoding the TCRm. 
     
     
         16 . The method of  claim 15 , wherein the TCRm comprises at least an antigen binding domain, a transmembrane domain, and an intracellular signaling domain; wherein optionally the antigen binding domain is capable of binding to a composite antigen comprising a peptide and a human leukocyte antigen (HLA) molecule; and wherein optionally the HLA molecule is a class I or class II HLA molecule. 
     
     
         17 . The method of  claim 16 , wherein the peptide comprises an alpha fetoprotein (AFP) peptide. 
     
     
         18 . The method of  claim 17 , wherein the composite antigen comprises an AFP peptide and a HLA-A2 molecule; and wherein the cancer is hepatocellular carcinoma. 
     
     
         19 . The method of  claim 16 , wherein the peptide comprises a preferentially expressed antigen in melanoma (PRAME) peptide; wherein optionally the PRAME peptide comprises SEQ ID NO: 41; wherein optionally the composite antigen comprises a PRAME peptide and a HLA-A*0201 molecule; and wherein the cancer is B-ALL, AML, multiple myeloma, T cell lymphoma, melanoma, non-small cell lung cancer, colon adenocarcinoma, or breast adenocarcinoma. 
     
     
         20 . The method of  claim 16 , wherein the peptide comprises a WT1 peptide and optionally a HLA-A2 molecule; and wherein the cancer is AML. 
     
     
         21 . The method of  claim 16 , wherein the antigen binding domain comprises an antibody or an antigen binding fragment thereof, or a non-antibody protein scaffold; and optionally wherein the antibody is a monoclonal antibody, a polyclonal antibody, a synthetic antibody, a human antibody, a humanized antibody, or a single domain antibody; optionally wherein the antigen binding fragment comprises a single chain variable fragment (scFv). 
     
     
         22 . The method of  claim 16 , wherein the intracellular signaling domain comprises a functional signaling domain of at least one stimulatory molecule; wherein optionally the at least one stimulatory molecule comprises a zeta chain associated with a T cell receptor complex or a CD3 zeta chain; and wherein optionally the intracellular signaling domain optionally further comprises a functional signaling domain of at least one costimulatory molecule. 
     
     
         23 . The method of  claim 22 , wherein the at least one costimulatory molecule comprises 4-1BB, CD28, CD27, CD134 (OX40), ICOS, DAP10, or DAP12. 
     
     
         24 . The method of  claim 6 , wherein the cells are further modified to comprise an exogenous cytokine, growth factor, antibody or antigen binding fragment, or any combination thereof, wherein the antibody or antigen binding fragment optionally comprises a bispecific T cell engager (BiTE). 
     
     
         25 . (canceled) 
     
     
         26 . (canceled) 
     
     
         27 . (canceled) 
     
     
         28 . A pharmaceutical composition comprising isolated CD3 + CD56 +  type III natural killer T cells, which are modified to express a CAR, TCR, or TCRm, and a pharmaceutically acceptable carrier. 
     
     
         29 . The pharmaceutical composition of  claim 28 , wherein the cells are CD3 + CD4 + CD56 + , CD3 + CD8 + CD56 + , or CD3 + CD4 − CD8 − CD56 +  cells, each optionally isolated from a biological sample of the subject or a donor. 
     
     
         30 . (canceled) 
     
     
         31 . (canceled) 
     
     
         32 . A method of preparing a therapy for treating or preventing a cancer in a subject in need thereof, comprising:
 a. isolating one or more CD3 + CD56 +  type III natural killer T cells from a biological sample; and   b. culturing the one or more CD3 + CD56 +  type III natural killer T cells in a growth medium to produce an expanded cell population.   
     
     
         33 . The method of  claim 32 , further comprising modifying the one or more CD3 + CD56 +  type III natural killer T cells to express a CAR, TCR, or TCRm, wherein the modifying comprises introducing one or more polynucleotides encoding the CAR, TCR, or TCRm into the one or more cells; wherein introducing one or more polynucleotides optionally comprises electroporation, transduction, and/or transfection; wherein optionally the one or more polynucleotides comprise mRNA and/or DNA; and wherein optionally the DNA comprises transposon DNA. 
     
     
         34 . The method of  claim 33 , wherein the one or more polynucleotides comprise one or more vectors, wherein the one or more vectors comprise one or more viral vectors or lentiviral vectors or γ-retroviral vectors. 
     
     
         35 . The method of  claim 32 , wherein the cancer is a solid tumor or a hematological malignancy; B-cell malignancy, leukemia, lymphoma, myeloma, or melanoma; acute myeloid leukemia (AML), B-cell precursor acute lymphoblastic leukemia (B-ALL), non-Hodgkin lymphoma (NHL), chronic lymphocytic leukemia (CLL), Ewing sarcoma, osteosarcoma, fibrosarcoma, rhabdomyosarcoma, mantle cell carcinoma, breast cancer or breast adenocarcinoma, lung adenocarcinoma, ovarian cancer, multiple myeloma, glioblastoma, hepatocellular cancer or hepatocellular carcinoma, neuroblastoma, metastatic melanoma, synovial sarcoma, bladder cancer, esophageal cancer, head and neck cancer, non-small cell lung cancer, prostate cancer, T cell lymphoma, or colon adenocarcinoma.